Investigating the Time-Dependent Reversibility of DCM-Induced Epigenetic, Matrix, and Functional Remodeling

研究 DCM 诱导的表观遗传、基质和功能重塑的时间依赖性可逆性

• 批准号: 10535260
• 负责人: Isabella Reichardt
• 金额: $ 4.16万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2025-09-15
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10535260
• 关键词: ATAC-seq; Aftercare; Age; Age-Months; Cardiac; Cardiac Myocytes; Cardiology; Cardiovascular Diseases; Cardiovascular system; Cells; Chromatin; Collagen; Congestive Heart Failure; Coupled; Cytoskeleton; DNA Methylation; Data; Deposition; Dilated Cardiomyopathy; Disease; Dose; Doxycycline; EFRAC; Engineering; Ensure; Epigenetic Process; Etiology; Extracellular Matrix; Family suidae; Fellowship; Fibroblasts; Fibrosis; Functional disorder; Future; Gene Expression; Generations; Genes; Genetic Transcription; Genome; Genomics; Goals; Heart; Heart Diseases; Heart Hypertrophy; Heart failure; Human; Hypertrophy; In Vitro; Induced Mutation; Inherited; Injury; Learning; Left ventricular structure; Link; Mechanics; Medicine; Memory; Mentors; Mentorship; Mesenchymal Stem Cells; Microfilaments; Modeling; Mus; Muscle Cells; Mutate; Mutation; Myocardial dysfunction; Myocardium; Myosin ATPase; Nature; Organ; Outcome; Patients; Periodicity; Pharmacological Treatment; Pharmacology; Phenotype; Physicians; Plant Roots; Point Mutation; Progressive Disease; Proteomics; Research; Resolution; Running; Scientist; Signal Transduction; Structure; Symptoms; Technology; Testing; Therapeutic; Time; Training; Transgenic Mice; Treatment Failure; Troponin C; United States National Institutes of Health; career; career development; chromatin remodeling; clinically relevant; effective therapy; epigenetic memory; epigenome; experimental study; familial dilated cardiomyopathy; healing; heart function; improved; inhibitor; mechanical stimulus; mouse model; multiple omics; mutant; palliative; pre-doctoral; pressure; professor; response; screening; therapeutic target; tool; transcriptome sequencing; transgene expression; translational therapeutics

Project Summary Dilated cardiomyopathy (DCM) is a highly prevalent inherited cardiac disease, characterized by systolic dysfunction, eccentric hypertrophy, and left ventricle dilation. While pharmacologic and mechanical treatments have been shown to partially improve cardiac function, these results are often short lived and highly variable from patient to patient. Moreover, recent studies have demonstrated that epigenetic and matrix dysregulation can persist, even in patients with improved systolic function after treatment. Given that aberrant chromatin remodeling and extracellular matrix (ECM) deposition have been identified as drivers of dilated remodeling—and that chromatin and ECM remodeling can become irreversible over time—it is crucial to understand the time- dependent effects of DCM mutations on reversing maladaptive remodeling at the genome, myocyte, and matrix levels. The central hypothesis of this proposal is that the reversibility of the DCM phenotype is time-dependent due to the accumulation of permanent ECM and chromatin remodeling as the disease progresses. To complete this proposal, I will utilize a DCM mouse model created by the Davis lab which contains a mutation (I61Q) in cardiac troponin C (cTnC) that directly decreases the myofilament’s Ca2+ sensitivity, leading to eccentric hypertrophy, systolic dysfunction, and left ventricle dilation. Importantly, expression of this mutant can be temporally controlled with doxycycline to specifically test our central hypothesis. In this proposal I will 1) Determine the time-dependent effects of I61Q cTnC expression on myocyte, matrix, and chromatin accessibility, 2) Examine the reversibility of DCM remodeling in myocyte, matrix, and chromatin accessibility at different stages of the disease, and 3) Determine if myocytes retain epigenetic memories of the mechanical disequilibrium caused by DCM. Improving our understanding of the time-dependent reversibility of DCM remodeling will better inform therapeutic targets and treatment windows for patients with DCM. Moreover, completion of this project will enhance Bella’s training as an independent scientist and prepare her to one day become a professor in cardiovascular engineering. Receiving the NIH F31 predoctoral fellowship will facilitate important experiments and training that will aid in her pursuit of this career goal. In this project, Bella will gain expertise in multi-omic approaches—such as proteomics, RNAseq, and ATACseq—which are growing in popularity due to their unbiased screening capabilities. The UW Genomics Core will assist Bella in learning how to effectively use these tools, which will not only benefit this project but will also be an incredibly useful skillset for Bella’s future career. Given the clinical relevance of this project, we have engaged Farid Moussavi-Harami, MD, an acting physician- scientist who practices cardiology within the UW Department of Medicine. Dr. Moussavi-Harami’s input as a clinician will be crucial for ensuring our research questions and aims remain relevant to patients, and his mentorship throughout this project will enhance Bella’s training and career development as she aims to eventually run a lab with an emphasis in translational therapeutics and technologies for cardiovascular diseases.

项目概要 扩张型心肌病（DCM）是一种高度流行的遗传性心脏病，其特征是收缩性心肌病 功能障碍、离心性肥大和左心室扩张。在药物和机械治疗的同时 已被证明可以部分改善心脏功能，但这些结果通常是短暂的且变化很大 从一个病人到另一个病人。此外，最近的研究表明表观遗传和基质失调 即使在治疗后收缩功能有所改善的患者中，这种情况也可以持续存在。鉴于染色质异常 重塑和细胞外基质 (ECM) 沉积已被确定为扩张重塑的驱动因素，并且 随着时间的推移，染色质和 ECM 重塑可能变得不可逆转——了解时间至关重要—— DCM 突变对逆转基因组、肌细胞和基质的适应不良重塑的依赖性影响 水平。该提案的中心假设是 DCM 表型的可逆性是时间依赖性的 由于随着疾病的进展，永久性 ECM 和染色质重塑的积累。完成 在这个提案中，我将利用 Davis 实验室创建的 DCM 小鼠模型，其中包含一个突变 (I61Q) 心肌肌钙蛋白 C (cTnC) 直接降低肌丝的 Ca2+ 敏感性，导致离心 肥厚、收缩功能障碍和左心室扩张。重要的是，该突变体的表达可以 用强力霉素进行时间控制，以专门测试我们的中心假设。在这个提案中我将 1) 确定 I61Q cTnC 表达对肌细胞、基质和染色质可及性的时间依赖性影响， 2) 检查不同阶段DCM重塑的肌细胞、基质和染色质可及性的可逆性 3) 确定肌细胞是否保留了引起的机械不平衡的表观遗传记忆 通过 DCM。提高我们对 DCM 重塑的时间依赖性可逆性的理解将更好地为我们提供信息 DCM 患者的治疗目标和治疗窗口。此外，该项目的完成将 加强贝拉作为一名独立科学家的培训，并为她有一天成为一名教授做好准备 心血管工程。获得 NIH F31 博士前奖学金将促进重要实验 以及有助于她追求这一职业目标的培训。在这个项目中，贝拉将获得多组学方面的专业知识 蛋白质组学、RNAseq 和 ATACseq 等方法由于其优点而越来越受欢迎 公正的筛选能力。华盛顿大学基因组学核心将帮助贝拉学习如何有效地使用这些 工具，这不仅有利于这个项目，而且对贝拉未来的职业生涯来说也是非常有用的技能。 鉴于该项目的临床相关性，我们聘请了代理医师 Farid Moussavi-Harami 医学博士 - 在华盛顿大学医学系从事心脏病学工作的科学家。 Moussavi-Harami 博士作为 临床医生对于确保我们的研究问题和目标与患者保持相关性至关重要，他的 整个项目的指导将加强贝拉的培训和职业发展，因为她的目标是 最终经营了一个实验室，重点研究心血管疾病的转化疗法和技术。